Analysis Tools
reproductive system
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• most testicular, caput and cauda epididymal spermatozoa display an additional ACRBP and PNA-positive punctuate structure associated with various aspects of the sperm acrosome and sperm head, unlike wild-type spermatozoa
• other material that is ACRBP/PNA-negative is associated with the concave surface of the sperm head, suggesting the presence of residual, possibly cytoplasmic, material that is not removed during spermiation
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• ACRBP/PNA positive material can be found near the apex of the sperm acrosome, near the top of the acrosomal cap or overlaying the acrosomal cap and equatorial segment, suggesting abnormal acrosome formation
• severely abnormal acrosomes are present, as shown by spermatozoa that exhibit only a diffuse staining for ACRBP and PNA; such spermatozoa have a sickle-shaped head but lack the pointed apex
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• presence of ACRBP/PNA-positive vesicle-like structure suggests that some proacrosomal vesicles may have not fused completely into a single acrosomal vesicle during the Golgi phase
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• mutant sperm undergo capacitation at a faster rate than wild-type sperm: time required for 50% of sperm to be capacitated, but remain acrosome intact, is 19 and 34 min, respectively
• at 30 min, ~70% of mutant sperm are of pattern B vs only ~ 28% of wild-type sperm; however, after 45 min, the level of capacitation reaches a similar plateau of 85%-90%, for sperm of either genotype
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• on a congenic C57BL/6J background, male homozygotes are severely subfertile relative to wild-type control males
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• at 9 hrs of incubation with cumulus-intact cells, no fertilized two-pronucleus egg is observed among eggs exposed to mutant sperm, whereas 40-51% of eggs incubated with wild-type sperm are fertilized
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• in vitro, the egg-binding ability of capacitated mutant sperm is only 50% that of wild-type sperm
• Background Sensitivity: ability of mutant sperm to bind to the zona pellucida and fertilize eggs in vitro is more severely impaired on a congenic C57BL/6J background than on a mixed background involving 129P2/OlaHsd and C57BL/6J
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• both testicular and epididymal spermatozoa show absence of ACRBP (proacrosin binding protein) proteolytic processing from its precursor to its mature form, unlike wild-type spermatozoa
• sperm extracts (containing only the precursor form of ACRBP) lack mature isoforms of acrosin, suggesting that autoactivation of proacrosin is less efficient
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• capacitated mutant sperm display increased sensitivity to zona-induced acrosomal exocytosis relative to wild-type sperm
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cellular
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• most testicular, caput and cauda epididymal spermatozoa display an additional ACRBP and PNA-positive punctuate structure associated with various aspects of the sperm acrosome and sperm head, unlike wild-type spermatozoa
• other material that is ACRBP/PNA-negative is associated with the concave surface of the sperm head, suggesting the presence of residual, possibly cytoplasmic, material that is not removed during spermiation
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• ACRBP/PNA positive material can be found near the apex of the sperm acrosome, near the top of the acrosomal cap or overlaying the acrosomal cap and equatorial segment, suggesting abnormal acrosome formation
• severely abnormal acrosomes are present, as shown by spermatozoa that exhibit only a diffuse staining for ACRBP and PNA; such spermatozoa have a sickle-shaped head but lack the pointed apex
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• presence of ACRBP/PNA-positive vesicle-like structure suggests that some proacrosomal vesicles may have not fused completely into a single acrosomal vesicle during the Golgi phase
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